Enzyme-linked immunosorbent assay for Shiga toxin and Shiga-like toxin II using P1 glycoprotein from hydatid cysts.
Shiga toxin from Shigella dysenteriae type 1 strains and Shiga-like toxins (SLT) I and II from Escherichia coli bind to terminal alpha-D-Galp-(1----4)-D-Galp containing glycolipids. Hydatid cyst fluid isolated from sheep infected with Echinococcus granulosus contains a glycoprotein (P1gp) with a terminal alpha-D-Galp-(1----4)-D-Galp disaccharide. Preparations of P1gp were shown to interact directly with Shiga toxin and to inhibit the binding and cytotoxicity of Shiga toxin to HeLa cells. A sandwich ELISA was developed using preparations of P1gp as the toxin capture molecule, which, with an appropriate polyclonal antibody, was capable of detecting as little as 80 pg/well Shiga toxin and 132 pg/well SLT-II. Thus, the P1gp-toxin interaction forms the basis for a simple antigen-capture ELISA that may be useful clinically for the rapid detection and quantitation of Shiga and Shiga-like toxins.
Acheson
D W
DW
Department of Medicine, New England Medical Center, Boston, MA 02111.
Keusch
G T
GT
Lightowlers
M
M
Donohue-Rolfe
A
A
D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis* / Q000276:immunology; D001667:Binding, Competitive; D001789:Blood Group Antigens / Q000276:immunology*; D002240:Carbohydrate Sequence; D004443:Echinococcosis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010127:P Blood-Group System / Q000276:immunology*; D022641:Shiga Toxin 2; D022603:Shiga Toxins
D001427:Bacterial Toxins; D001789:Blood Group Antigens; D006023:Glycoproteins; D010127:P Blood-Group System; D022641:Shiga Toxin 2; D022603:Shiga Toxins